Detection of Rickettsia in ticks using Loop-mediated Isothermal Amplification (LAMP)

Article

Lansdell, S., Hassan, M. M., La Ragione, R., Betson, M., Núncio, M. S., de Carvalho, I. S., Zé-Zé, L., de Sousa, R. and Cutler, S. 2025. Detection of Rickettsia in ticks using Loop-mediated Isothermal Amplification (LAMP). CMI Communications. p. In press. https://doi.org/10.1016/j.cmicom.2025.105069
AuthorsLansdell, S., Hassan, M. M., La Ragione, R., Betson, M., Núncio, M. S., de Carvalho, I. S., Zé-Zé, L., de Sousa, R. and Cutler, S.
Abstract

Objectives
The objective of the study was to develop a Loop-mediated Isothermal Amplification (LAMP) assay for screening of Rickettsia species circulating in ticks using the citrate synthase gene (gltA). The LAMP assay employed portable visualisation methods, making the assay more field-suitable. Furthermore, prior methods have not used gltA as the target, despite proven success in Polymerase Chain Reaction (PCR) methods.

Methods
Using an alignment of 72 DNA sequences (comprised of 21 Rickettsia species) from GenBank we designed a novel set of gltA LAMP primers. Evaluation used DNA from 12 Rickettsia species as positive controls (extracted from cultures or naturally infected ticks) alongside a panel of negative controls representing different bacterial species. Subsequently this assay was used to screen 295 Ixodes ricinus and 24 I. hexagonus ticks collected from the UK (including northern and southern England and northern Scotland).

Results
LAMP successfully detected 11 out of 12 (91.7%) Rickettsia species, excluding Rickettsia akari. Among 319 ticks collected in the UK, three were positive for Rickettsia (0.9%). All three positives were I. ricinus ticks, while none of the 24 I. hexagonus ticks were positive. Results were confirmed using a published PCR method. Sanger sequencing of PCR amplicons generated for each positive tick showed that they were all R. helvetica.

Conclusions
This study introduces a novel field-applicable LAMP protocol for efficient Rickettsia screening in ticks to better assess its prevalence and consequent health risks. Furthermore, this assay has proven suitability for rickettsial detection in I. ricinus ticks, which has been reported as unsuccessful in previous European studies.

Graphical abstract
Created in BioRender. Cutler, S. (2025) https://BioRender.com/m74g606

JournalCMI Communications
Journal citationp. In press
ISSN2950-5909
Year2025
PublisherElsevier
Publisher's version
License
File Access Level
Anyone
Digital Object Identifier (DOI)https://doi.org/10.1016/j.cmicom.2025.105069
Publication dates
Online06 Mar 2025
Publication process dates
Accepted26 Feb 2025
Deposited07 Mar 2025
Copyright holder© 2025 The Authors
Permalink -

https://repository.uel.ac.uk/item/8z22z

Log in to edit

Download files

Publisher's version
1-s2.0-S2950590925000113-main.pdf
License: CC BY-NC-ND 4.0
File access level: Anyone

  • 2
    total views
  • 0
    total downloads
  • 2
    views this month
  • 0
    downloads this month

Export as

Related outputs

Insights into the contribution of multiple factors on Ixodes ricinus abundance across Europe spanning 20 years using different machine learning algorithms
Lansdell, S., Zorto, A., Seto, M., Negera, E., Sharif, S. and Cutler, S. 2025. Insights into the contribution of multiple factors on Ixodes ricinus abundance across Europe spanning 20 years using different machine learning algorithms. Ticks and Tick-borne Diseases. 16 (1), p. Art. 102437. https://doi.org/10.1016/j.ttbdis.2025.102437
Machine Learning-Based Techniques for Assessing Critical Factors for European Tick Abundance
Zorto, A., Lansdell, S., Seto, M., Negera, E., Sharif, S. and Cutler, S. 2025. Machine Learning-Based Techniques for Assessing Critical Factors for European Tick Abundance. International Journal of Computer Theory and Engineering. 17 (1), pp. 13-20.
Innovative Ensemble Approaches for Assessing Critical Factors for European Tick Abundance
Zorto, A., Lansdell, S., Seto, M., Gobena, E., Cutler, S. and Sharif, S. 2024. Innovative Ensemble Approaches for Assessing Critical Factors for European Tick Abundance. 2024 International Conference on Innovation and Intelligence for Informatics, Computing, and Technologies. IEEE.
Comparative Analysis of Tick-Borne Relapsing Fever Spirochaetes from Ethiopia and Nigeria
Bankole, A. A., Kumsa, B., Mamo, G., Ogo, N. I., Elelu, N., Morgan, W. and Cutler, S. J. 2023. Comparative Analysis of Tick-Borne Relapsing Fever Spirochaetes from Ethiopia and Nigeria. Pathogens. 12 (Art. 81). https://doi.org/10.3390/pathogens12010081
Coxiella burnetii
Cutler, S. 2023. Coxiella burnetii. in: Food Science Elsevier.
Novel approaches for the serodiagnosis of louse-borne relapsing fever
Röttgerding, F., Njeru, J., Schlüfter, E., Latz, A.., Mahdavi, R., Steinhoff, U., Cutler, S. J., Besier, S., Kempf, V. A. J., Fingerle, V. and Kraiczy, P. 2022. Novel approaches for the serodiagnosis of louse-borne relapsing fever. Frontiers in Cellular and Infection Microbiology. 12 (Art. 983770). https://doi.org/10.3389/fcimb.2022.983770
Borrelia duttonii-like spirochetes parasitizec Meriones persicus in East Azerbaijan Province of Iran
Ghasemi, A., Naddaf, S. R., Mahmoudi, A., Rohani, M., Naeimi, S., Mordadi, A., Cutler, S. and Mostafavi, E. 2021. Borrelia duttonii-like spirochetes parasitizec Meriones persicus in East Azerbaijan Province of Iran. Ticks and Tick-borne Diseases. 12 (Art. 101825). https://doi.org/10.1016/j.ttbdis.2021.101825
Tick-borne diseases and co-infection: Current considerations
Cutler, S. J., Vayssier-Taussat, M., Estrada-Peña, A., Potkonjak, A., Mihalca, A. D. and Zeller, H. 2020. Tick-borne diseases and co-infection: Current considerations. Ticks and Tick-borne Diseases. 12 (Art. 101607). https://doi.org/10.1016/j.ttbdis.2020.101607
Serospatial epidemiology of zoonotic Coxiella burnetii in a cross section of cattle and small ruminants in northern Nigeria
Elelu, N., Bankole, A., Musa, R. J., Odetokun, I. A., Rabiu, M., Biobaku, K. T., Aremu, A., Ahmed, A. O., Ghali, M. I., Raji, M. A., Ogo, N. I., Cutler, S. and Ogundipe, G. A. T. 2020. Serospatial epidemiology of zoonotic Coxiella burnetii in a cross section of cattle and small ruminants in northern Nigeria. PLoS ONE. 15 (Art. e0240249). https://doi.org/10.1371/journal.pone.0240249
Rejection of the name Borreliella gen. nov. and all proposed species comb. nov. placed therein. Request for an opinion
Margos, G., Stevenson, B., Gofton, A., Oskam, C., Estrada-Pena, A., Cutler, S. and Fingerle, V. 2020. Rejection of the name Borreliella gen. nov. and all proposed species comb. nov. placed therein. Request for an opinion. International Journal of Systematic and Evolutionary Microbiology. 70 (5), p. 3577–3581. https://doi.org/10.1099/ijsem.0.004149
Bartonella vinsonii sub. arupensis infection in animals of veterinary importance, ticks and biopsy samples
Chochlakis, D., Cutler, S., Giadinis, N. D. and Psaroulaki, A. 2020. Bartonella vinsonii sub. arupensis infection in animals of veterinary importance, ticks and biopsy samples. New Microbes and New Infections. 34 (Art. 100652). https://doi.org/10.1016/j.nmni.2020.100652
A review on the eco-epidemiology and clinical management of human granulocytic anaplasmosis and its agent in Europe
Matei, I. A., Estrada-Peña, A., Cutler, S., Vayssier-Taussat, M., Varela-Castro, L., Potkonjak, A., Zeller, H. and Mihalca, A. D. 2019. A review on the eco-epidemiology and clinical management of human granulocytic anaplasmosis and its agent in Europe. Parasites and Vectors. 12 (Art. 599). https://doi.org/10.1186/s13071-019-3852-6
A new Borrelia on the block: Borrelia miyamotoi – a human health risk?
Cutler, S., Vayssier-Taussat, Muriel, Estrada-Peña, Agustín, Potkonjak, Aleksandar, Mihalca, Andrei Daniel and Zeller, Hervé 2019. A new Borrelia on the block: Borrelia miyamotoi – a human health risk? Eurosurveillance. 24 (18), p. Art. 1800170. https://doi.org/10.2807/1560-7917.ES.2019.24.18.1800170
An updated meta-analysis of the distribution and prevalence of Borrelia burgdorferi s.l. in ticks in Europe
Estrada-Peña, Agustín, Cutler, S., Potkonjak, Aleksandar, Vassier-Tussaut, Muriel, Van Bortel, Wim, Zeller, Hervé, Rui, Natalia Fernández and Mihalca, Andrei Daniel 2018. An updated meta-analysis of the distribution and prevalence of Borrelia burgdorferi s.l. in ticks in Europe. International Journal of Health Geographics. 17, p. Art. 41. https://doi.org/10.1186/s12942-018-0163-7
Ornithodoros savingyi – the tick vector of Candidatus Borrelia kalaharica in Nigeria
Cutler, S., Idris, Jibrin M., Ahmed, Akeem O. and Elelu, Nusirat 2018. Ornithodoros savingyi – the tick vector of Candidatus Borrelia kalaharica in Nigeria. Journal of Clinical Microbiology. 56 (9), pp. e00532-18. https://doi.org/10.1128/JCM.00532-18
Migrant health—a cause for concern?
Cutler, S. and Bottieau, E. 2017. Migrant health—a cause for concern? Clinical Microbiology and Infection. 23 (5), pp. 281-282. https://doi.org/10.1016/j.cmi.2017.04.014
Adding a further twist to the tail of Leptospirosis in the UK
Cutler, S. 2017. Adding a further twist to the tail of Leptospirosis in the UK. Veterinary Record. 180 (17), p. Editorial. https://doi.org/10.1136/vr.j1985
Critical Aspects for Detection of Coxiella burnetii
Mori, Marcella, Mertens, Katja, Cutler, S. and Santos, Ana Sofia 2017. Critical Aspects for Detection of Coxiella burnetii. Vector-Borne and Zoonotic Diseases. 17 (1), pp. 33-41. https://doi.org/10.1089/vbz.2016.1958
Diagnosing Borreliosis
Cutler, S., Rudenko, Nataliia, Golovchenko, Maryna, Cramaro, Wibke J., Kirpach, Josiane, Savic, Sara, Christova, Iva and Amaro, Ana 2017. Diagnosing Borreliosis. Vector-Borne and Zoonotic Diseases. 17 (1), pp. 2-11. https://doi.org/10.1089/vbz.2016.1962
Relapsing fever causative agent in southern Iran is a closely related species to East African borreliae
Naddaf, Saied Reza, Ghazinezhad, Behnaz, Kazemirad, Elham and Cutler, S. 2017. Relapsing fever causative agent in southern Iran is a closely related species to East African borreliae. Ticks and Tick-borne Diseases. 8 (6), pp. 882-886. https://doi.org/10.1016/j.ttbdis.2017.07.006
Emerging borreliae – Expanding beyond Lyme borreliosis
Cutler, S., Ruzic-Sabljic, Eva and Potkonjack, Aleksandar 2016. Emerging borreliae – Expanding beyond Lyme borreliosis. Molecular and Cellular Probes. 31, pp. 22-27. https://doi.org/10.1016/j.mcp.2016.08.003
A Case of Canine Borreliosis in Iran Caused by Borrelia persica
Shirani, Darush, Rakhshanpoor, Alaleh, Cutler, S., Ghazinezhad, Behnaz and Naddaf, Saied Reza 2016. A Case of Canine Borreliosis in Iran Caused by Borrelia persica. Ticks and Tick-borne Diseases. 7 (3), pp. 424-426. https://doi.org/10.1016/j.ttbdis.2015.12.020
Evolution of Koch's postulates: towards a twenty-first century understanding of microbial infection
Antonelli, Guido and Cutler, S. 2016. Evolution of Koch's postulates: towards a twenty-first century understanding of microbial infection. Clinical Microbiology and Infection. 22 (7), pp. 583-584. https://doi.org/10.1016/j.cmi.2016.03.030
There is inadequate evidence to support the division of the genus Borrelia
Margos, Gabriele, Marosevic, Durdica, Cutler, S., Derdakova, Marketa, Diuk-Wasser, Maria, Emler, Stefan, Fish, Durland, Gray, Jeremy, Hunfeld, Klaus-Peter, Jaulhac, Benoit, Kahl, Olaf, Kovalev, Sergey, Kraiczy, Peter, Lane, Robert S., Lienhard, Reto, Lindgren, Per-Eric, Ogden, Nicholas, Ornstein, Katharina, Rupprecht, Tobias, Schwartz, Ira, Sing, Andreas, Straubinger, Reinhard K., Strle, Franc, Voordouw, Maarten, Rizzoli, Annapaola, Stevenson, Brian and Fingerle, Volker 2016. There is inadequate evidence to support the division of the genus Borrelia. International Journal of Systematic and Evolutionary Microbiology. 67 (4), pp. 1081-1084. https://doi.org/10.1099/ijsem.0.001717
Emerging horizons for tick-borne pathogens: from the ‘one pathogen–one disease’ vision to the pathobiome paradigm
Vayssier-Taussat, Muriel, Kazimirova, Maria, Hubalek, Zdenek, Hornok, Sándor, Farkas, Robert, Cosson, Jean-François, Bonnet, Sarah, Vourch, Gwenaël, Gasqui, Patrick, Mihalca, Andrei Daniel, Plantard, Olivier, Silaghi, Cornelia, Cutler, S. and Rizzoli, Annapaola 2015. Emerging horizons for tick-borne pathogens: from the ‘one pathogen–one disease’ vision to the pathobiome paradigm. Future Microbiology. 10 (12), pp. 2033-2043.
Refugee crisis and re-emergence of forgotten infections in Europe
Cutler, S. 2015. Refugee crisis and re-emergence of forgotten infections in Europe. Clinical Microbiology and Infection. 22 (1), pp. 8-9.
Relapsing Fever Borreliae: A Global Review
Cutler, S. 2015. Relapsing Fever Borreliae: A Global Review. Clinics in Laboratory Medicine. 35 (4), pp. 847-865.
Neglected zoonoses: forgotten infections among disregarded populations
Cutler, S. 2015. Neglected zoonoses: forgotten infections among disregarded populations. Clinical Microbiology and Infection. 21 (5), pp. 392-393.
Urban Leptospirosis in Africa: A Cross-Sectional Survey of Leptospira Infection in Rodents in the Kibera Urban Settlement, Nairobi, Kenya
Halliday, J. E. B., Knobel, D. L., Allan, K. J., de C. Bronsvoort, B. M., Handel, I., Agwanda, B., Cutler, S., Olack, B., Ahmed, A., Hartskeerl, R. A., Njenga, M. K., Cleaveland, S. and Breiman, R. F. 2013. Urban Leptospirosis in Africa: A Cross-Sectional Survey of Leptospira Infection in Rodents in the Kibera Urban Settlement, Nairobi, Kenya. American Journal of Tropical Medicine and Hygiene. 89 (6), pp. 1095-1102.
New concepts for the old challenge of African relapsing fever borreliosis
Cutler, S., Abdissa, A. and Trape, J.-F. 2009. New concepts for the old challenge of African relapsing fever borreliosis. Clinical Microbiology and Infection. 15 (5), pp. 400-406.
Myths, legends and realities of relapsing fever borreliosis
Cutler, S. 2009. Myths, legends and realities of relapsing fever borreliosis. Clinical Microbiology and Infection. 15 (5), pp. 395-396.
Does RecA have a role in Borrelia recurrentis?
Cutler, S., Rinky, I. J. and Bonilla, E. M. 2011. Does RecA have a role in Borrelia recurrentis? Clinical Microbiology and Infection. 17 (2), pp. 195-197.
Spirochaetes: past lessons to future directions [Editorial]
Cutler, S. 2011. Spirochaetes: past lessons to future directions [Editorial]. Clinical Microbiology and Infection. 17 (4), pp. 481-483.
The polymicrobial nature of biofilm infection
Wolcott, R., Costerton, J. W., Raoult, D. and Cutler, S. 2012. The polymicrobial nature of biofilm infection. Clinical Microbiology and Infection. 19 (2), pp. 107-112.
Dangerous liaisons-our expanding appreciation of microbial synergism [Editorial]
Cutler, S. 2013. Dangerous liaisons-our expanding appreciation of microbial synergism [Editorial]. Clinical Microbiology and Infection. 19 (2), pp. 105-106.
Relapsing fever borreliosis
Cutler, S. 2015. Relapsing fever borreliosis. in: Schlossberg, David (ed.) Clinical Infectious Disease Cambridge Cambridge University Press. pp. 1068-1071
High Prevalence of Rickettsia africae Variants in Amblyomma variegatum Ticks from Domestic Mammals in Rural Western Kenya: Implications for Human Health
Maina, Alice N., Jiang, Ju, Omulo, Sylvia A., Cutler, S., Ade, Fredrick, Ogola, Eric, Feikin, Daniel R., Njenga, M. Kariuki, Cleaveland, Sarah, Mpoke, Solomon, Ng'ang'a, Zipporah, Breiman, Robert F., Knobel, Darryn L. and Richards, Allen L. 2014. High Prevalence of Rickettsia africae Variants in Amblyomma variegatum Ticks from Domestic Mammals in Rural Western Kenya: Implications for Human Health. Vector-Borne and Zoonotic Diseases. 14 (10), pp. 693-702.
Portuguese Hosts for Ornithodoros erraticus Ticks
Palma, Mariana, Lopes de Carvalho, Isabel, Osório, Hugo, Zé-Zé, Líbia, Cutler, S. and Núncio, Maria Sofia 2013. Portuguese Hosts for Ornithodoros erraticus Ticks. Vector-Borne and Zoonotic Diseases. 13 (10), pp. 775-777.
Urban Leptospirosis in Africa: A Cross-Sectional Survey of Leptospira Infection in Rodents in the Kibera Urban Settlement, Nairobi, Kenya
Halliday, J. E. B., Knobel, D. L., Allan, K. J., de C. Bronsvoort, B. M., Handel, I., Agwanda, B., Cutler, S., Olack, B., Ahmed, A., Hartskeerl, R. A., Njenga, M. K., Cleaveland, S. and Breiman, R. F. 2013. Urban Leptospirosis in Africa: A Cross-Sectional Survey of Leptospira Infection in Rodents in the Kibera Urban Settlement, Nairobi, Kenya. American Journal of Tropical Medicine and Hygiene. 89 (6), pp. 1095-1102.
Fibroblast growth stimulation, DPPH antioxidant assay and antimicrobial activities of Funtumia elastica (Preuss) Stapf (Apocynaceae) leaf extracts
Osei-Djarbeng, Samuel N., Cutler, S., Cutler, Ronald R. and Corcoran, O. 2014. Fibroblast growth stimulation, DPPH antioxidant assay and antimicrobial activities of Funtumia elastica (Preuss) Stapf (Apocynaceae) leaf extracts. European Journal of Medicinal Plants. 4 (7), pp. 835-843. https://doi.org/10.9734/EJMP/2014/9316
Brucellosis in Ethiopia
Yohannes, Moti, Degefu, Hailu, Tolosa, Tadele, Belihu, Kelay, Cutler, Ronald R. and Cutler, S. 2013. Brucellosis in Ethiopia. African Journal of Microbiology Research. 7 (14), pp. 1150-1157.
Antimicrobial and anti-quorum sensing activities of some Ghanaian medicinal plants
Osei-Djarbeng, S., Corcoran, O., Cutler, S. and Cutler, R. R. 2012. Antimicrobial and anti-quorum sensing activities of some Ghanaian medicinal plants. International Journal of Indigenous Medicinal Plants. 28 (10), pp. 221-226.
Typing African Relapsing Fever Spirochetes
Scott, Julie Christine, Wright, David Julian Maurice and Cutler, S. 2005. Typing African Relapsing Fever Spirochetes. Emerging Infectious Diseases. 11 (11), pp. 1722-1729.
Possibilities for Relapsing Fever Reemergence
Cutler, S. 2006. Possibilities for Relapsing Fever Reemergence. Emerging Infectious Diseases. 12 (3), pp. 369-374.
Population Structure of East African Relapsing Fever Borrelia spp.
Cutler, S., Bonilla, E. Margarita and Singh, Rajbir J. 2010. Population Structure of East African Relapsing Fever Borrelia spp. Emerging Infectious Diseases. 16 (7), pp. 1076-1080.
Human Complement Regulators C4b-Binding Protein and C1 Esterase Inhibitor Interact with a Novel Outer Surface Protein of Borrelia recurrentis
Grosskinsky, Sonja, Schott, Melanie, Brenner, Christiane, Cutler, S., Simon, Markus M. and Wallich, Reinhard 2010. Human Complement Regulators C4b-Binding Protein and C1 Esterase Inhibitor Interact with a Novel Outer Surface Protein of Borrelia recurrentis. PLoS Neglected Tropical Diseases. 4 (6).
Public Health Threat of New, Reemerging, and Neglected Zoonoses in the Industrialized World
Cutler, S., Fooks, Anthony R. and van der Poel, Wim H.M. 2010. Public Health Threat of New, Reemerging, and Neglected Zoonoses in the Industrialized World. Emerging Infectious Diseases. 16 (1), pp. 1-7.
The Genome of Borrelia recurrentis, the Agent of Deadly Louse-Borne Relapsing Fever, Is a Degraded Subset of Tick-Borne Borrelia duttonii
Lescot, Magali, Audic, Stéphane, Robert, Catherine, Nguyen, Thi Tien, Blanc, Guillaume, Cutler, S., Wincker, Patrick and Couloux, Arnaud 2008. The Genome of Borrelia recurrentis, the Agent of Deadly Louse-Borne Relapsing Fever, Is a Degraded Subset of Tick-Borne Borrelia duttonii. PLoS Genetics. 4 (9), pp. 1-11. https://doi.org/10.1371/journal.pgen.1000185
Borrelia recurrentis Employs a Novel Multifunctional Surface Protein with Anti-Complement, Anti-Opsonic and Invasive Potential to Escape Innate Immunity
Grosskinsky, Sonja, Schott, Melanie, Brenner, Christiane, Cutler, S., Kraiczy, Peter, Zipfel, Peter F., Simon, Markus M. and Wallich, Reinhard 2009. Borrelia recurrentis Employs a Novel Multifunctional Surface Protein with Anti-Complement, Anti-Opsonic and Invasive Potential to Escape Innate Immunity. PLoS ONE. 4 (3), pp. 1-12.